Oil spills can cause tremendous environmental damage and are a serious threat to public health. Petroleum contains many toxic compounds such as polyaromatic hydrocarbons (PAHs). Some PAH compounds are highly carcinogenic and cause DNA mutations in humans. When petroleum is spilled into the sea, PAH compounds rapidly spread through the marine environment and eventually accumulate in marine life and threaten our seafood supply. The FDA has mandated that petroleum-contaminated seafood is a human health threat and should not be harvested or sold. Therefore, after an oil spill, careful monitoring is needed to assess the extent of the contaminated areas, ensure that seafood supplies are free of PAH compounds and monitor the return of affected areas to a normal state to allow re-opening of closed areas. Unfortunately, despite this critical need, the most currently used testing method has been shown to be somewhat inaccurate and unreliable for the detection of potentially hazardous levels of PAH compounds in seafood. New alternative testing methods are needed. Our proposed research will create improved tests to detect petroleum components in seafood. We will develop new enzyme-based tests to detect PAH compounds in seafood using dioxygenases enzymes from microbes. These enzymes will be used to generate a color change when PAH compounds are present in the sample. Our simple enzyme tests can be directly used in microplates, a widely-accepted, inexpensive, high-throughput format for food testing applications. To ease the transition to our new enzymatic format, our assays will be designed to work well with currently accepted seafood processing methods. We will also investigate ways to shorten the sample processing steps to increase the throughput-capacity of the test. Our new test kits will provide seafood producers and government agencies such as the FDA and NOAA with much-needed cost-effective, reliable and sensitive tools to detect petroleum in seafood after oil spills. PUBLIC HEALTH RELEVANCE: Oil spills are major threats to public health and the environment. Petroleum contains many toxic and carcinogenic substances;these substances can get into seafood near oil spill sites. It is important to test for toxic oil substances in seafood after an oil spill, but current methods are not adequate for thorough testing. Our proposed research will produce novel enzyme-based tests for oil contamination in seafood. The improved reliable properties of our new method will greatly assist the FDA to ensure seafood safety.